Inhibition of tumor growth by antisense oligonucleotides for IL-8 and IL-8 receptor

ABSTRACT

Oligonucleotides are provided which are effective in inhibiting the growth, metastasis and/or angiogenesis of tumors, including particularly melanoma and/or lung cancer. Methods are also provided for use of these oligonucleotides in the treatment of diseases.

This application is a divisional of application Ser. No. 08/796,031filed Feb. 5, 1997, now U.S. Pat. No. 5,849,903 which is a CIP ofapplication Ser. No. 08/561,302 filed Nov. 21, 1995 (abandoned).

I. FIELD OF THE INVENTION

The field of the invention is oligonucleotide therapeutics, and moreparticularly the use of oligonucleotides to modulate the expression ofIL-8 and/or IL-8 Receptor to control growth, metastasis and/orangiogenesis in tumors.

II. BACKGROUND OF THE INVENTION IL-8

Interleukin-8 (IL-8, neutrophil activating protein-1, or NAP-1) is amember of C-X-C chemokine family of related cytokines having broadinvolvement in inflammatory responses, tissue injury, growth regulationand cellular adhesion. Cerretti, D. P., et al., MolecularCharacterization of Receptors for Human Interleukin-8, GRO/MelanomaGrowth-Stimulatory Activity and Neutrophil Activating Peptide-2,Molecular Immunology, 30(4), 359-367 (1993); and Koch, A. E., et al., Insitu expression of cytokines and cellular adhesion molecules in the skinof patients with systemic sclerosis, Pathobiology, 61(5-6), 239-46(1993). A review of the C-X-C family is given in Streiker, R. M., etal., Role of C-X-C Chemokines As Regulators Of Angiogenesis In LungCancer, J. of Leukocyte Biology, 57, 752-762 (1995). IL-8 has also beenshown to have a potent stimulatory effect on angiogenesis. See, e.g.,Koch, A. E., Interleukin-8 as a Macrophage-Derived Mediator ofAngiogenesis, Science, 258, 1798-1800 (1992).

It is known that IL-8 is produced by a variety of normal human somaticcells including monocytes/macrophages, dermal fibroblasts, vascularendothelial cells, keratinocytes, and mesangeal cells. Yasumoto, K., etal., Tumor Necrosis Factor Alpha and Interferon Gamma SynergisticallyInduce Interleukin 8 Production in a Human Gastric Cancer Cell LineThough Acting Concurrently on AP-1 and NF-kB-like Binding Sites of theInterleukin 8 Gene, J. of Biological Chemistry, 267(31), 22506-11(1992). Apparently, such cells produce IL-8 only when stressed, and notunder conditions of normal growth and homeostasis,. Factors which induceIL-8 production include inflammation, IL-1, TNF, LPS and thrombin.

It is also known that Il-8 is commonly secreted by tumor cells. Tumor isherein defined to mean uncontrolled and progressive multiplication ofcells, including benign tumors and all forms of neoplasm and cancer. Inone study IL-8 was shown to be present in 80% of carcinoma cell linesand 83.3% of cancer tissues studied. It has also been shown that IL-8 isa growth factor for cancers, and that anti-IL-8 antibody and IL-8receptor anti-sense are each be effective in attenuating cancer cellgrowth in vitro. Ishoko, T., et al, Supra; Schadendorf, D., et al., IL-8Produced by Human Malignant Melanoma Cells in Vitro Is an EssentialAutocrine Growth Factor, J. Immunol., 151(5), 2667-2674 (1993).

Because of its effects on growth, it is suspected that IL-8 has asignificant role in the metastatic spread of melanoma and other cancers.Schadendorf, D., et al., Supra. It has been demonstrated, for example,that IL-8 is directly involved in the mechanism of melanoma metastasis,(Singh, R. K., et al, Ultraviolet B Irradiation Promotes Tumorigenic andMetastatic Properties in Primary Cutaneous Melanoma via Induction ofInterleukin 8, Cancer Res., 55, 3669-3674, 1995), as well as lungcancer, (Greenspoon, S. A., et al., The expression of IL-8 duringTumorgenesis and Metastasis in a model of human nonsmall cell lungcancer/SCID mouse chimera, Clinical Research, 42, 404A, 1994). IL-8 hasalso been found to play role in tumor neovascularization, for example,in head and neck squamous cell carcinomas. Cohen R. F., et al, ArchOtolaryngol. Head Neck Surg., 121, 202-209, 1995. Thus, IL-8 playsimportant role in tumor growth, metastasis and angiogenesis.

IL-8 RECEPTOR

IL-8 is a ligand for cell-membrane IL-8 Receptor, and it is thought thatinteraction between IL-8 and IL-8 Receptor is required for IL-8 action.Two IL-8 receptor genes have been identified so far, IL-8 Receptor typeA and type B. Both genes belong to the so-called seven transmembranedomain, G protein-coupled receptor family. Receptor A has been shown tobe activated by IL-8, and receptor B has been shown to be activated byIL-8 as well as other cytokines belonging to C-X-C family includingMelanoma Growth Stimulatory Activity (MGSA). Receptor B is said to beremarkable for its "promiscuity," binding with growth regulatorygene(GRO)(also known as melanoma growth stimulatory activity (MGSA)),neutrophil activating peptide-2 (NAP-2), and neutrophil activatingpeptide-1 (CAP-1), all of which cluster on human chromosome 4q13-q21.

Ahuja, S. K., Molecular Evolution of the Human Interleukin-8 ReceptorGene Cluster, Nature Genics, 2, 31-36 (1992); Cerretti, et al., supra.Common effects elicited by IL-8 and MGSA on neutrophils include calciummobilization, induction of respiratory burst, degranulation of secretoryvesicles, and an increase in both GTPase activity and GTP binding.

IL-8 Receptor B has been found not only in neutrophils and other cellsof hematopoietic lineage, but also in non-hematopoietic cells. IL-8Receptor B has been found, for example, in cells responsive to MGSA,tumor necrosis factor and tumor promoters including placenta and kidneycell lines. Mueller, S. G., et al, Melanoma Growth Stimulatory ActivityEnhances the Phosphorylation of the Class II Interleukin-8 Receptor inNon-hematopoietic Cells, J. Biol. Chem., 269, 1973-1980 (1994); NF-κBSubunit-Specific Regulation of the Interleukin-8 Promoter, Mol. & Cell.Bio., 13(10) 6137-6146 (1993). The role and function of IL-8 Receptor Bpresent in cancer and other tumor cells is not fully elucidated. Thereis, however, evidence that activation of IL-8R B (1) is involved in themechanism of growth regulation of melanoma and tumorigenic fibroblasts;(2) is associated with transformation of lung cells by asbestos, and (3)correlates with metastic potential of melanoma.

cDNAs coding for at least two IL-8 receptors have been isolated andassigned to chromosome 2q35. Morris, S. W., et al., Assignment of theGenes Encoding Human Interleukin-8 Receptor Types 1 and 2 and anInterleukin-8 Receptor Pseudogene to Chromosome 2q 35, Genomics, 14,685-691 (1992); Murphy, P. M., and Tiffany, H. L., Cloning ofComplementary DNA Encoding a Functional Huiman Interleukin-8 Receptor,Science, 253, 1280-1283 (1991). Two additional IL-8 receptors have alsobeen identified biochemically, and may correspond to the cDNA'spreviously identified. Moser, et al., Expression of Transcripts for TwoInterleukin Receptors in Human Phagocytes, Lymphocytes and MelanomaCells, Biochem., 294, 285-292 (1993).

Given the growth stimulatory effect of IL-8 on cells responsive tovarious tumor growth factors, it would be advantageous to provideantisense oligonucleotides which modulate expression of either IL8 orIL-8 Receptor in cancers in vivo. It would be particularly advantageousto provide oligonucleotides which are effective against lung cancer andmelanoma because each of these cancers produce their own growth factors.

There are at least two major types of lung cancer, small cell lungcarcinoma (SCLC) and non-small cell lung carcinoma (NSCLC). SCLCcomprises approximately one-fourth of the cases, expressesneuroendocrine markers, and generally metastasizes early to lymph nodes,brain, bones, lung and liver. NSCLC comprises the majority of theremaining lung tumor types, and includes adeno-carcinoma, squamous cellcarcinoma, and large cell carcinoma. NSCLC is characterized byepithelial-like growth factors and receptors, and is locally invasive.

Melanoma cells, unlike normal melanocytes, can proliferate in theabsence of exogenous growth factors. This independence apparentlyreflects the production of growth factor and cytokines kines forautocrine growth stimulation, including TGF-Å, TGF-, platelet-derivedgrowth factor A and B chains, basic fibroblast growth factor, IL-8,IL-6, IL-1, granulocyte macrophage colony stimulating factor, and MGSA.Guo Y, et al., Inhibition of Human Melanoma Growth and Metastasis inVivo by Anti-CD44 Monolclonal Antibody. Cancer Res., 54, 1561-1565(1994).

To date some success has been reported in antisense oligonucleotides ata 20 oM concentration to reduce the expression of IL-8 Receptor andinhibit the growth of lung cancer cells (Ishiko, T., et al, Supra), andmalignant melanoma cells (Schadendorf, D., et al, Supra). These reports,however, reflected in vitro studies, and it is not at all clear that thesame oligonucleotides would be effective in vivo. Even with respect toin vitro work, at least one researcher reported success only withmodified phosphorothioate anti-sense oligonucleotides. Bargou, R. C.,Regulation of Proliferation In The New Hodgkin Cel Line HD-MYZ byAntisense-Oligonucleotides Against IL-6 and IL-8, Molecular Basis ofCancer Therapy, edited by E. J. Freidreich, S. A. Stass (1995)Cambridge, Mass., by Blackwell Science Inc. Thus, there is still a needfor oligonucleotides which inhibit the growth, metastasis and/orangiogenesis of melanoma and lung cancer in vivo through interferenewith the expression of IL-8 and/or IL-8 Receptor. Moreover, since Il-8and IL-Receptors appear to be involved generally in the mechanisms oftumor growth, metastasis and angiogenesis, phenomena which arecharacteristic of most types of cancer, there is a need to target one orgenes in treating cancers and other tumors in general.

III. SUMMARY OF THE INVENTION

Oligonucleotides are provided which are effective in treatment oftumors, particularly melanoma, lung, prostate and breast cancer. Methodsare also provided for use of these oligonucleotides in the treatment ofdiseases.

IV. BRIEF DESCRIPTION OF THE DRAWINGS

The novel features believed characteristic of the invention are setforth and differentiated in the appended claims. The invention inaccordance with preferred and exemplary embodiments, together withfurther advantages thereof, is more particularly described in thefollowing detailed description taken in conjunction with theaccompanying drawing in which:

FIG. 1 is a bar graph showing growth-inhibitory effect of controlOligonucleotide ICN 131, IL-8 Oligonucleotide ICN-70 (SEQ ID NO:2) andIL-8 Oligonucleotide ICN-133 (SEQ ID NO:3) at concentration 0.1 μM onmelanoma cells (ATCC HTB140), lung carcinoma (ATCC HTB 177), prostatecancer (ATCC HTB81) and breast cancer cells (T47D) during 72 hrs oftreatment. The oligonucleotide designations are as follows:

    ICN-131 CAC CCG AAG GAA CC  (SEQ ID NO:1)                                     ICN-70  GAA AGT TTG TGC CTT ATG GA                                                                        (SEQ ID NO:2)                                     ICN-133 GGC GCA GTG TGG TCC (SEQ ID NO:3)                                 

FIG. 2 is a bar graph showing the growth-inhibitory effect ofIL-Receptor B Oligonucleotide ICN-247 (SEQ ID NO:5), ICN-107 (SEQ IDNO:4) and ICN-297 (SEQ ID NO:6) at concentration 0.1 μM on melanoma cell(ATCC HTB140), non-small lung carcinoma(ATCC HTB177), prostate carcinoma(ATCC HTB81), and breast cancer cells (T47D) during 72 hrs treatment.The oligonucleotide designations are as follows:

    ICN-107 CTT CCT GTC TGA TGG CTT CT                                                                        (SEQ ID NO:4)                                     ICN-247 TGA AAA TTT AGA TCA TC                                                                            (SEQ ID NO:5)                                     ICN-297 AGT GTG CGT GTG G   (SEQ ID NO:6)                                 

FIG. 3 is a table showing the kinetic of subcutaneous melanoma growth inbalb/c nude/nude mice expressed as slope following linear regressionanalysis for untreated mice, and mice treated with ICN 131 and ICN-70(SEQ ID NO:2).

FIG. 4 is a table showing the kinetic of subcutaneous lung cancer growthin balb/c nude/nude mice expressed as slope following linear regressionanalysis for untreated mice, and mice treated with ICN 131 and ICN-70(SEQ ID NO:2).

V. DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS

Synthesis of Oligonucleotides

The discussion below describes the production of various oligonucleotidesequences, and the testing of the sequences for their effectivenessagainst lung and/or melanoma cancer cells. All tested sequences weresynthesized on an automated DNA synthesizer (Applied Biosystems model394) as phosphodiester oligonucleotides using standard phosphodiesterchemistry. The resulting oligonucleotides were purified by HPLC using areverse phase semiprep C8 column (ABI) with linear gradient of 5%acetonitrile in 0.1 M triethylammonium acetate and acetonitrile. Thepurity of the products was checked by HPLC using an analytical C18column (Beckman).

In Vitro Testing

Testing of the oligonucleotide sequences was carried out on large celllung carcinoma line ATCC HTB 177, melanoma cancer cells ATCC HTB 140,prostate cancer ATCC HTB81 and breast cancer cells T47D. The cells wereroutinely propagated in culture in a humidified incubator at 37° C. in5% CO₂ / 95% air at atmospheric pressure. Both cell lines were grown asadherent cultures in 90% RPMI 1640, supplemented with 10% fetal bovineserum. Cells were seeded in 96-well microtiter plates at 2,000 cells perwell, and subsequently treated once with 0.1 μM concentration of theantisense or control oligonucleotides. After 72 h of culture, all cellswere pulsed with 1 μCi/well/200 μl of [³ H] thymidine, cultured for anadditional 2 h, harvested with an automatic Harvester 96 (Tomtec), and[³ H] thymidine incorporation determined in a β-counter. To assureaccuracy of the results, all cultures were performed in triplicate andrepeated at least three times. The results are presented at FIG. 1 andFIG. 2

In Vivo Testing

The in vivo experiments summarized in FIGS. 3 and 4 were carried out inBalb/c Nude/Nude mice deficient in T lymphocytes but not deficient in Blymphocytes or NK cells. According to the protocol, 2.5 million ofmelanoma cells (ATCC HBT140) were injected subcutanously inHanks-balanced salt solution. 10 days later tumor size was measured, andthe same day oligonucleotides were injected intratumoraly at aconcentration 0.1 μM in 100 μL of Hanks- balanced salt Solution.Injection was repeated twice a day for 14 days. Tumor size mm² wasmeasured 7, 10, and 14 days of treatment. At last day of treatment (day14) animals were sacrificed for collection of tumor tissues and fortheir analysis on the histopathological level and gene/proteinexpression level. In case of lung cancer cells, 0.25 million of thecells were injected (ATCC HBT 177). Subsequently, animals were treatedas described for melanoma.

Pharmaceutical Preparations of Oligonucleotides

As with other oligonucleotides, the oligonucleotides of the presentinvention may be adapted for administration to the body in a number ofways suitable for the selected method of administration, includingorally, intravenously, intramuscularly, intraperitoneally, topically,and the like. In addition to comprising one or more differentoligonucleotides, the subject pharmaceutical formulations may compriseone or more non-biologically active compounds, i.e., excipients, such asstabilizers (to promote long term storage), emulsifiers, binding agents,thickening agents, salts, preservatives, and the like.

Delivery of oligonucleotides as described herein is well known in theart for a wide range of animals, including mammals, and especiallyincluding humans. For example, Delivery Strategies for AntisenseOligonucleotide Therapeutics, CRC press (Saghir Akhtar, ed. 1995)details man)y such delivery routes and strategies, and is incorporatedherein by reference in its entirety. By way of example only, and withoutlimiting the applicability of the entire book, chapter describesadministration by traditional intravenous, intraperitoneal andsubcutaneous routes, along with "non-damaging routes" such asintranasal, ocular, transdermal and iontophoresis routes (Id. at 71-83),all of which are applicable to the present invention. Chapter 6 of thesame text deals with modifications to make oligonucleotide therapeutics,nuclease resistant, and the terms nucleotide(s), oligonucleotide(s) andnucleic acid base(s) as used herein specifically includes the describedmodifications and all other conservatively modified variants of thenatural form of such compounds. (Id. at 85-104). The claimedoligonucleotides can also be bonded to a lipid or other compound that isactively transported across a cell membrane, either with or without alinker, and administered orally as disclosed in U.S. Pat. No. 5,411,947,which is also incorporated herein by reference. Still further, theoligonucleotides of the present invention can be administered in a"naked" form, encapsulated, in association with vesicles, liposomes,beads, microspheres, as conjugates, and as an aerosol directly to thelung, using for example ICN Biomedicals product no. SPAG 2. Thus,oligonucleotides of the present invention can be administeredsubstantially by all known routes of administration foroligonucleotides, using all accepted modifications to produce nucleotideanalogs and prodrugs, and including all appropriate binders andexcipients, dosage forms and treatment regimens.

Oligonucleotides can be employed in dosages and amounts which areconventional in the art for the underlying bio-active compound, butadjusted for more efficient absorption, transport and cellular uptake.The dosages may be administered all at once, or may be divided into anumber of smaller doses which are then administered at varying intervalsof time. At present the most preferred administration of theoligonucleotides of the present invention comprises intravenousadministration of between about 0.1 and 100 mg of oligonucleotide per kgof body weight of the patient, 1-14 times per week for approximately 40days. This regimen is based upon the observed half-life for similaroligonucleotides in vivo of several minutes to several hours, along withthe observation that the effect upon protein synthesis may last up to 48or 72 hours. The specific treatment regimen given to any individualpatient will, of course, depend upon the experience of the clinician inweighing the disease involved, the health and responsiveness of thepatient, side effects, and many other factors as is well known amongsuch clinicians. For example, greater of lesser dosage levels, andtreatment regimens covering greater or lesser periods of time would bedependent upon the judgment of the attending clinician, and may includeperiods of rest during which treatment with the oligonucleotide istemporarily halted. Treatment may also be combined with other anticancerand palliative treatments as appropriate. Progression/remission of thedisease being treated may be determined by following tumor size, extentof metastasis and other factors through radiological analysis and othermeans known in the art, and the existence and extent of side effects maybe determined by following functioning of the liver and/or kidneys, andby following the blood circulation as for example through the use ofEKG, which again are well known in the art.

Oligonucleotides may be administered in any convenient manner, such asby oral, intravenous, intraperitoneal, intramuscular, or subcutaneous orother known routes. However, for many of these routes, modifiedoligonucleotides, including backbone and/or sugar modified nucleotidesas set forth in U.S. patent application Ser. No. 08/333,895 and08/333,545, respectively, the disclosures of which are incorporatedherein by reference, may be used advantageously to enhance survivabilityof the oligonucleotides. For oral administration, oligonucleotides maybe administered with an inert diluent or with an assimilable ediblecarrier, or oligonucleotides may be incorporated directly with the foodof the diet. Orally administered oligonucleotides may be incorporateswith excipients and used in the form of ingestible tablets, buccaltablets, troches, capsules, elixirs, suspension syrups, wafers, and thelike.

The tablets, troches, pills, capsules and the like may also contain thefollowing: a binder, such as gum tragacanth, acacia, cornstarch, orgelatin; excipients, such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agents, such assucrose, lactose or saccharin; a flavoring agent such as peppermint, oilof wintergreen, or cherry flavoring. When the dosage unit is a capsule,it may contain, in addition to materials of the above type, a liquidcarrier. Various other materials may also be present as coatings or tootherwise modify the physical form of the dosage unit. For instance,tablets, pills, or capsules may be coated with shellac, sugar or both. Asyrup or elixir may contain sucrose as a sweetening agent, methyl andpropylparabens as preservatives, a dye and flavoring such as cherry orrange flavor. Such additional materials should be substantiallynon-toxic in the amounts employed. Furthermore, the oligonucleotides maybe incorporated into sustained-release preparations and formulations.

Formulations for parenteral administration may include sterile aqueoussolutions or dispersions, and sterile powders for the extemporaneouspreparation of sterile, injectable solutions or dispersions. Thesolutions or dispersions may also contain buffers, diluents, and othersuitable additives, and may be designed to promote the cellular uptakeof the oligonucleotides in the composition, e.g., the oligonucleotidesmay be encapsulated in suitable liposomes. Preferably the solutions anddispersions for parenteral administration are sterile and sufficientlyfluid for proper administration, sufficiently stable from manufactureand use, and preserved against the contaminating action ofmicroorganisms such as bacteria and fungi. The carrier can be a solventor dispersion medium containing, for example, water, ethanol, polyol(for example, glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), suitable mixtures thereof, and vegetable oils.The proper fluidity can be maintained, for example, by the use of acoating such as lecithin, by maintenance of the required particle sizein the case of dispersion and by the use of surfactants.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with one ormore of the various other ingredients described above, followed bysterilization. Dispersions may generally be prepared by incorporatingthe various sterilized active ingredients into a sterile vehicle whichcontains the basic dispersion medium and the required other ingredientsfrom those listed above. In the case of sterile powders used to preparesterile injectable solutions, the preferred methods of preparation arevacuum-drying and freeze-drying techniques which yield a powder of theactive ingredient plus any additional desired ingredient from previouslysterile-filtered solutions.

Pharmaceutical formulations for topical administration may be especiallyuseful with certain bio-active compounds for localized treatment.Formulations for topical treatment included ointments, sprays, gels,suspensions, lotions, creams, and the like. Formulations for topicaladministration may include, in addition to the subject oligonucleotides,known carrier materials such as isopropanol, glycerol, paraffin, stearylalcohol, polyethylene glycol, etc. The pharmaceutically acceptablecarrier may also include a known chemical absorption promoter. Examplesof absorption promoters are e.g., dimethylacetamide (U.S. Pat. No.3,472,931), trichloro-ethanol or trifluoroethanol (U.S. Pat. No.3,891,757), certain alcohols and mixtures thereof (British Patent No.1,001,949), and British patent specification No. 1,464,975.

Solutions of the oligonucleotides may be stored and/or administered asfree base or pharmacologically acceptable salts, and may advantageouslybe prepared in water suitably mixed with a surfactant such ashydroxypropylcellulose. Dispersions can also be prepares in glycerol,liquid polyethylene glycols, and mixtures thereof and in oils. Thesecompositions and preparations may advantageously contain a preservativeto prevent the growth of microorganisms.

The prevention of the action of microorganisms can be brought about byvarious antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thmerosal, and the like. In manycases, it will be preferable to include isotonic agents such as sodiumchloride. Prolonged of injectable compositions can be brought about bythe use of agents which delay absorption, such as aluminum monostearateand gelatin.

The compositions and preparations described preferably contain at least0.1% of active oligonucleotide. The percentage of the compositions andpreparations may, of course, be varied, and may contain between about 2%and 60% of the weight of the amount administered. The amount of activecompounds in such therapeutically useful compositions and preparationsis such that a suitable dosage will be obtained.

As used herein, "pharmaceutically acceptable carrier" includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents and the like. The use ofsuch media and agents for pharmaceutical active substances is well knownin the art. Except insofar as any conventional media or agent isincompatible with the therapeutic active ingredients, its use in thetherapeutic compositions and preparations is contemplated. Supplementaryactive ingredients can also be incorporated into the compositions andpreparations.

In addition to the therapeutic uses of the subject oligonucleotides, theoligonucleotides may also be used in the laboratory tool for the studyof absorption, distribution, cellular uptake, and efficacy.

INCORPORATION BY REFERENCE

All patents, patents applications, and publications cited areincorporated herein by reference.

EQUIVALENTS

Thus, antisense oligonucleotides effective in controlling IL-8 Receptorexpression in non-small cell lung cancer and melanoma cells, have beendisclosed. Procedures for using these oligonucleotides in clinicalpractice have also been disclosed. While specific embodiments andapplications have been shown and described, it would be apparent tothose skilled in the art that additional modifications are possiblewithout departing from the inventive concepts herein. The invention,therefore, is not to be restricted except in the spirit of the appendedclaims.

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What is claimed is:
 1. A method for treating a cancer which is at leastpartially mediated by IL-8 Receptor, said method comprising the step ofadministering to a patient an effective amount of the followingoligonucleotide: 5' CTT CCT GTC TGA TGG CTT CT 3' (SEQ ID NO: 4).
 2. Amethod for treating a cancer which is at least partially mediated byIL-8, said method comprising the step of administering to a patient aneffective amount of an oligonucleotide having the sequence 5' GAA AGTTTG TGC CTT ATG GA 3' (SEQ ID NO:2).